DNA Plasmid Isolation Using Alkaline Lysis Method

DNA Plasmid Isolation Using Alkaline Lysis Method Buffers and Solutions Alkaline Lysis solution I: 50 mM glucose, 25 mM Tris-Cl (pH ), 10 mM EDTA (pH ), de-ion water Alkaline Lysis solution II : N NaOH, 1% (w/v) SDS, de-ion water Alkaline Lysis solution III : 5 M potassium acetate, glacial acetic acid, de-ion water Ethanol 70% (v/v) Isopropanol TE-RNAase pH Method 1. Pour overnight grown culture to mL labeled falcon tube. 2. Centrifugate at rpm for 1 min. 3. Remove the supernatant from the tube. 4. Repeat step 1-3, until leaves bacterial pellet as dry as possible. 5. Add 150 L resuspension buffer , resuspend the bacterial pellet properly by vortexing. 6. Add 200 L Lysis solution to bacterial suspension (freshly made), close the tube tightly and mix contents thoroughly by inverting the tube 4-6 times until the solution becomes viscious.

5. Add 150 µL resuspension buffer, resuspend the bacterial pellet properly by vortexing. 6. Add 200 µL lysis solution to bacterial suspension (freshly made), close the tube tightly and mix contents thoroughly by inverting the tube 4-6 times until the solution becomes viscious. 7.

Tags:

  Buffer, Lysis

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